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Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
Published on: January 17, 2015
Isolation and characterization of anti ROR1 single chain fragment variable antibodies using phage display technique
Leili Aghebati-Maleki1,2,3, Vahid Younesi4,5, Farhad Jadidi-Niaragh2,6
1Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Abstract:
Receptor tyrosine kinase-like orphan receptor (ROR1) belongs to one of the families of receptor tyrosine kinases (RTKs). RTKs are involved in the various physiologic cellular functions including proliferation, migration, survival, signaling and differentiation. Several RTKs are deregulated in various cancers implying the targeting potential of these molecules in cancer therapy. ROR1 has recently been shown to be expressed in various types of cancer cells but not in normal adult cells. Hence a molecular inhibitor of extracellular domain of ROR1 that inhibits ROR1-cell surface interaction is of great therapeutic importance. In an attempt to develop molecular inhibitors of ROR1, we screened single chain variable fragment (scFv) phage display libraries, Tomlinson I + J, against one specific synthetic oligopeptide from extracellular domain of ROR1 and selected scFvs were characterized using various immunological techniques. Several ROR1 specific scFvs were selected following five rounds of panning procedure. The scFvs showed specific binding to ROR1 using immunological techniques. Our results demonstrate successful isolation and characterization of specific ROR1 scFvs that may have great therapeutic potential in cancer immunotherapy.
Insights
Researchers developed specific single chain variable fragments (scFvs) targeting Receptor Tyrosine Kinase-Like Orphan Receptor 1 (ROR1). These ROR1-targeting scFvs show potential for cancer immunotherapy development.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- Receptor tyrosine kinases (RTKs) regulate crucial cellular functions.
- Dysregulated RTKs are implicated in various cancers.
- Receptor tyrosine kinase-like orphan receptor 1 (ROR1) is expressed in cancer cells but not normal adult cells, making it a therapeutic target.
Purpose of the Study:
- To develop molecular inhibitors targeting the extracellular domain of ROR1.
- To identify and characterize single chain variable fragments (scFvs) that bind specifically to ROR1.
- To explore the therapeutic potential of ROR1-specific scFvs in cancer immunotherapy.
Main Methods:
- Screening of Tomlinson I + J single chain variable fragment (scFv) phage display libraries.
- Panning against a synthetic oligopeptide from the extracellular domain of ROR1.
- Characterization of selected scFvs using immunological techniques.
Main Results:
- Isolation of several ROR1-specific scFvs after five rounds of panning.
- Demonstrated specific binding of isolated scFvs to ROR1 via immunological assays.
- Successful isolation and characterization of scFvs with potential therapeutic applications.
Conclusions:
- Specific ROR1-targeting scFvs have been successfully developed.
- These scFvs exhibit specific binding to ROR1.
- The identified ROR1 scFvs hold significant therapeutic promise for cancer immunotherapy.
